Enzymes

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Enzymes

ISBN 9789394681521

Publication Date Mon Aug 01 00:00:00 UTC 2011

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Table of contents

Biographical note

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This book Enzymes presents a broad and comprehensive analysis of Enzyme Technology with a distinct emphasison Enzyme Nomenclature, Enzyme biology, Enzyme Engineering and Biosensors. Written is lucid and student-friendly approach, this book is rich in theory and practice.

Cover
Title Page
Copyright Page
Contents
Preface
1 INTRODUCTION TO ENZYMES
- Introduction
- Meaning and Definition
- History
- General Properties of Enzymes
  - 1. Higher Reaction Rates
  - 2. Milder Reaction Condition
  - 3. Greater Reaction Specificity
  - 4. Capacity for Regulations
  - 5. Presence of Species Specificity
  - 6. Variations in Activity and Ability
  - 7. Substrate Specificity
  - 8. Activation and Inhibition
- Sources of Enzymes
- Characteristics of Enzymes
- Questions
2 ENZYME NOMENCLATURE AND CLASSIFICATION
- Nomenclature
- Classification
- Major Classes
  - 1. Oxidoreductases
  - 2. Transferases
  - 3. Hydrolases
  - 4. Lyases
  - 5. Isomerases
  - 6. Ligases (Synthetases)
- Key to Numbering and Classification of Enzymes
  - 1. Oxidoreductases
  - 2. Transferases
  - 3. Hydrolases
  - 4. Lyases
  - 5. Isomerases
  - 6. Ligases
- Examples
- Questions
3 ENZYME EXTRACTION AND PURIFICATION
- Importance of Enzyme Purification
- General Procedure
- Extraction
- Enzyme Histochemistry
- Use of Centrifugation
- Extraction of Soluble Enzymes
- Extraction of Membrane-Bound Enzymes
- Nature of the Extraction Medium
- Purification of Enzymes
  - Preliminary Procedure
  - Fractionation Methods
  - Partition Chromatography
  - Electrophoresis
  - Cation Exchange Chromatography
  - Gel Filteration
  - Affinity Chromatography
  - HPLC Technology
  - Lyophilization
- Determination of Molecular Weight of Enzymes
- Examples of Purification Procedure
- Adenylate Kinase from Pig Muscles
  - Ribulose Bisphosphate Carboxylase from Spinach
  - RNA Polymerase from E.Coli
  - Arom Multi Enzyme Protein from Neurospora
  - Glutathione Reductase from E.Coli
  - Adenylate Cyclase
  - Chymosin
- Questions
4 ENZYME ASSAY
- Factors Controlling in Assays
  - 1. Salt Concentration
  - 2. Temperature
  - 3. pH
  - 4. Substrate Saturation
  - 5. Level of Crowding
- Discontinous Assay
- Continous Assay
  - Coupled Assay
- Enzyme Assay Techniques
  - Visible and Ultraviolet Spectrophotometer Methods
  - Spectrofluorimetric Method
  - Luminescence Method
  - Radioisotope Method
- Questions
5 TYPES OF ENZYME
- Introduction
- Coenzymes
  - Characteristics of Coenzymes
- Isoenzymes
  - Lactate dehydrogenase enzyme (LDH)
- Monomeric Enzymes
  - Proteases (proteolytic enzymes)
- Oligomeric Enzymes
  - Lactate Dehydrogenase
  - Lactose Synthase
  - Tryptophan Synthase
  - Pyruvate Dehydrogenase
- Metalloenzymes and Metal-Activated Enzymes
  - Metalloenzymes
  - Metal Activation
- Extremozymes
- Abzymes
- Ribozymes
- Synzymes
- Bi and Polyfunctional Enzyme
- Questions
6 ENZYME CATALYSIS
- Mechanisms in Organic Chemistry
- Mechanisms of Enzyme Catalysis
  - Acid–Base Catalysis
  - Strain Distortion and Conformational Change
  - Catalysis by Approximation (Entrophic Contribution)
  - Covalent Catalysis(Nucleophilic vs Electrophilic Catalysis)
- Coenzymes in Enzyme-Catalyzed Reactions
- Questions
7 ENZYME SPECIFICITY
- Group Specificity
- Absolute Specificity
- Stereochemical Specificity
- The Active Sites
- Fischer’s Lock and Key Hypothesis
- Koshland Induced Fit Hypothesis
- Non-Productive Binding
- Transition State Stabilization
  - Cryoenzymology
- Questions
8 ENZYME KINETICS
- Goals of Enzyme Kinetics
- Chemical Kinetics
  - Reaction Order
  - Classification of Chemical Reactions by Kinetic Order
- Kinetics of Bisubstrate Enzyme Reactions
  - The Michaelis–Menten Equation
  - The Lineweaver–Burk Plot
  - The Eadie - Hofstee and Hans Plot
- Kinetics of Multisubstrate Enzyme-Catalyzed Reactions
  - Ping-Pong Bi-Bi Mechanism
  - Random-Order Mechanism
  - Compulsory-Order Mechanism
  - Steady-State Kinetics
  - Plots For Mechanisms Which Follow the General Rate Equation
- Allosteric Enzymes
  - Monod–Wyman Changeux (MWC) Model
  - MWC Model and Allosteric Regulation
- Questions
9 ENZYME INHIBITION
- Reversible Inhibition
  - Competitive Inhibition
  - Uncompetitive Inhibition
  - Non-competitive Inhibition and Mixed Inhibition
  - Partial Inhibition
  - Substrate Inhibition
  - Allosteric Inhibiton
- Irreversible Inhibition
- Questions
10 ENZYME COFACTORS
- Mode of Action
- Cofactors Acting as Carriers
  - Redox Carriers
  - Phosphate Carriers
  - CO2 Carrier
  - Amino Group Carriers
  - Acyl Group Carriers
  - Carrier of One Carbon Group
  - Sulphate Carrier
  - Aldehyde Carrier
- Questions
11 ENZYME BIOLOGY
- Introduction
- Intracelluar Localization of Enzymes
- Histochemical Methods
  - Separation of Particulate Fraction
- Questions
12 ENZYME ENGINEERING
- Definition
- Objectives
- Principle of Enzyme Engineering
- Procedure in Enzyme Engineering
- Example
- Protein Engineering
  - Addition of Disulphide Bond
  - Changing Asparagine to other Aminoacids
  - Reduction of Number of Free Sulphydryl Residues
  - Increase of Enzymatic Activity
  - Modification of Enzyme Specificity
- Secretion of Cloned Proteins
  - Large Scale Fermentation
  - Principles of Microbial Growth
  - Bioreactors
  - Harvesting of Microbial Cells
  - Disruption of Microbial Cells
  - Downstream Processing
- Questions
13 IMMOBILIZATION OF ENZYME
- Advantages
- Methods of Enzyme Immobilization
  - Adsorption
  - Covalent Bonding
  - Entrapment
  - Crosslinking
  - Encapsulation
- Applications of Immobilized Enzyme
- Immobilized Enzyme Kinetics
  - Heterogenous Concentrations
  - Concentration Gradients and Reaction Rates in Solid Catalyst
  - Effect of External Mass Transfer Resistance
  - Diffusion Effects in Surface-Bound Enzymes on Non-Porous Support Materials
- Design of Immobilized Enzyme Reactors
  - Batch Reactors
  - Continuous Flow Reactors
  - Packed Bed Reactor
- Fluidized-Bed Reactors
- Membrane Reactors
- Questions
14 BIOSENSORS
- Principle
- General Features
- Glucose Electrode
- Types of Biosensors
  - Calorimetric Biosensors
  - Amperometric Biosensors
  - Optical Biosensors
  - Acoustic Wave Biosensor
  - Potentiometric Biosensors
  - Whole Cell Biosensors (Microbial Biosensors)
- Applications of Biosensors
- Questions
15 CLINICAL ENZYMOLOGY
- Amylase
  - Clinical Significance
- Transaminases
  - Clinical Significance
- Alkaline Phosphatase (ALP)
  - Clinical Significance
- Acid Phosphatase (ACP)
  - Clinical Significance
- Lactate Dehydrogenase (LD)
  - Clinical Significance
- Creatine Kinase (CK)
  - Clinical Significance
- Gamma-Glutamyl Transferase (GGT)
  - Clinical Significance
- Cholinesterases
  - Clinical Significance
- Questions
16 APPLICATIONS IN ENZYMES
- Applications in Medicine
  - Assay of Plasma Enzymes
  - Therapeutic Uses
- Application In Industry
  - In Food Industry
  - Baking of Bread
  - Enzymes and Recombinant DNA Technology
  - In Dairy Industry
  - Cheese Making
  - In Starch Industry
  - In Detergent Industry
  - In Leather Industry
  - Wool Industry
  - Production Syrups
  - In Sugar Industry
  - In Brewing Industry
- Biotechnological Applications of Enzymes
- Analytical Uses
  - Principles of Enzymatic Analysis
- Questions

Dr. Devika R is Assistant Professor at NSS Training College, Ottapalam, Palghat. Her areas of specialisation include Mathematics, Education, ICT. She completed her Masters in Science from University of Kerala and completed her Masters in Education from Manonmaniam Sundaranar University and her doctorate from University of Kerala in 2011. She has over 12 years of teaching experience and has published more than 15 research papers in nationally reputed journals and contributed 2 articles in edited books.

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